Methods of improving central nervous system functioning

ABSTRACT

Methods of causing an improvement in central nervous system function are provided. The methods include administering an aliquot of stem cells to the patient, the cells being derived from blood, e.g., umbilical cord blood. In some cases a growth factor is administered with the cells.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of, and claims priority from, U.S.patent application Ser. No. 09/698,893, filed Oct. 27, 2000 (allowed),which is incorporated herein by reference in its entirety.

TECHNICAL FIELD

This invention relates to methods of improving central nervous systemfunctioning.

BACKGROUND

The central nervous system is a complex system of tissues, includingcells, fluids and chemicals that interact in concert to enable a widevariety of functions, including movement, navigation, cognition, speech,vision and emotion. Unfortunately, a variety of debilitatingmalfunctions of the central nervous system may occur and disrupt some orall of these functions. These malfunctions are broad in range andinclude, for example, missing genetic elements, e.g., genetic diseasessuch as Tay-Sachs; malfunctioning cellular processes, e.g., Parkinson'sDisease; trauma, e.g., head injury; degenerative diseases, e.g.,Alzheimer's Disease; and damage due to lack of oxygen, e.g., caused bystroke or asphyxiation.

Typically, treatments for restoring central nervous system functionafter damage by the above malfunctions have been limited to drugs, andto adaptive or behavioral therapies. These approaches are generallylimited in their ability to reverse damage (or stop degeneration) andrestore normal function.

Recent research has explored the possibility of using cells to restorefunction to the central nervous system. Data using animal modelssuggests that cell implantation or transplantation may be an effectivemeans for restoring central nervous system function after damage. Cellsthat have been used in this research have included variousnonhematopoietic precursor cells, for example, fetal or embryonic neuralcells from porcine and human sources (see, e.g., Nairne, S. P.,Animal-to-Human transplants; the ethics of Xenotransplantation. London:Nuffield Council of Bioethics, 1996); immortalized fetal neural cells(Ren et al., Co-Administration of Neural Stem Cells and bFGF EnhancesFunctional Recovery Following Focal Cerebral Infarction in Rat, Soc.Neurosci. Abstr., 26:2291, 2000); mesenchymal bone marrow stem andprogenitor cells (Chen et al., Intracerebral Transplantation withCultured Bone Marrow Stroma Cells after MCAO in Rats, American Societyfor Neural Transplantation & Repair, Program and Abstracts, Volume 7:2000, A-3) including multipotent adult stem cells (see, e.g., Keene etal., Phenotypic Expression of Transplanted Human Bone Marrow-DerivedMultipotent Adult Stem Cells into the Rat CNS, American Society forNeural Transplantation & Repair, Program and Abstracts, Volume 7: 2000,6-3); murine neural stem cells (Marciniak, Neural Stem Cells, InCombination with Basic FibroBlast Growth Factor (bFGF) May Represent aTreatment/or Stroke, supra, A-I), including immortalized murineneuroepithelial stem cells (Modo et al., Implantation Site of Stem Cellsin MCAD Rats Influences the Recovery on Different Behavioral Tests,supra, 8-2); adult mouse and human neural stem and progenitor cells(see, e.g., Steindler et al., Stem Cells in the Human Brain, supra,8-3); fetal mesencephalic cells (Mendez et al., SimultaneousIntraputaminal and Instranigral Fetal Dopaminergic Grafts in Parkinson'sDisease: First Clinical Trials, supra, 5-3); testis-derived Sertolicells (Cameron et al., Formation o/SNACs by Simulated MicrogravityCoculture of Sertoli Cells and NT2 Cells. supra, C-), and crude bonemarrow extract (Mahmood et al., Effects of Transplantation of BoneMarrow Cells on Wistar Rats Following Traumatic Brain Injury, supra,A-4). To overcome the lack of availability of many of these types ofcells, researchers have even resorted to studying the possibility ofadministering cancer cells such as teratacarcinomal cells (Kondziolka etal., Transplantation of Cultured Human Neuronal Cells/or Patients withStroke, Neurology 2000, 55: 565-569), despite the inherent dangers ofthe use of cancerous cells. Some research into cellular therapies hasreached the clinical stage. Generally, the cells that have been used inthe research described above pose potential hazards to patients, and/orare difficult to obtain.

SUMMARY

The present invention features methods of enhancing recovery of centralnervous system function by administering cells derived from blood, e.g.,cells derived from cord blood. By “cord blood”, we mean blood that isderived from the placenta or umbilical cord around the time of the birthof a human infant. Advantageously, there is a readily available supplyof such cells, without resort to fetal or embryonic sources or toharvesting the patient's own brain cells. For example, cord blood cellsare currently banked for autologous or allogeneic administration inother applications. As a result, these cells need not be immortalized,and therefore there is a reduced risk of possible cancerous outgrowthsor other detrimental complications that may result from the use ofimmortalized cells. Because these cells are primary cells, they also areless likely to be carcinogenic or cause other related problems. Also,the use of autologous or immunologically matched cord blood cellsreduces the risk of rejection.

Moreover, in some cases, e.g., if a large supply of cord blood is notreadily available, a suitable cell population can be obtained using arelatively small sample of cord blood or other source of relativelyundifferentiated precursor cells, e.g., as described in U.S. Pat. Nos.5,674,750 and 5,925,567, the entire disclosures of which areincorporated herein by reference.

A method of causing an improvement in function of the central nervoussystem of a subject, comprising administering to the subject an aliquotof cells derived from umbilical cord blood.

In one aspect, the invention features a method of causing an improvementin a function of the central nervous system of a subject, comprisingadministering to the subject an aliquot of cells derived from blood, thealiquot containing stem cells.

In another aspect, the invention features a method of causing animprovement in a function of the central nervous system of a subject,comprising administering to the subject an aliquot of cells derived fromblood and a growth factor.

Implementations of the invention may include one or more of thefollowing features. The cells are derived from umbilical cord blood. Thecells are derived from peripheral blood. The method further includesobtaining the aliquot of cells by separating a desired cell populationfrom the cord blood. The growth factor is selected from the groupconsisting of oncostatin M and growth factors from the followingfamilies: FGF, neurotrophin, IGF, CNTF, EGF, TGF-beta, LIF,interleukins, PDGF and VEGF. The method further includes obtaining asample of cells and purifying the sample to obtain the aliquot. Themethod further includes obtaining a sample of cells and expanding atleast a selected population of cells in the sample ex vivo to obtain thealiquot. The aliquot of cells includes allogeneic cells. Alternatively,or in addition, the aliquot of cells includes autologous cells. Theimprovement results in recovery from a central nervous system trauma,repair of central nervous system damage or disease, and/or regenerationof central nervous system tissue. The improvement includes measurablestroke recovery. The improvement is the result of stroke repair. Theimprovement results from tissue regeneration after a stroke. Theimprovement results from a genetic element contained in the administeredcells. The genetic element is endogenous to the administered cells. Thegenetic element has been exogenously added to the administered cells.The improvement includes head trauma recovery and/or repair. Theimprovement results from tissue regeneration after head trauma. Thecells are administered intercerebrally. The cells are CD 34+/−, Lin−cells or precursor cells. The cells are characterized as: CD2⁻, CD3⁻,CD14⁻, CD16⁻, CD19⁻, CD24⁻, CD56⁻, CD66b⁻, glycophorin A⁻, flk-1⁺,CD45⁺, CXCR4⁺, MDR⁺. The improvement results from treatment of one ofthe following diseases: Parkinson's Disease, Alzheimer's Disease,Huntington's Disease, ALS, MS, Tay-Sacks, and cerebral palsy. The methodfurther includes administering to the subject a cell differentiationfactor or a neural guidance molecule.

In a further aspect, the invention features a method of causing animprovement in central nervous system function of a patient including(i) obtaining an aliquot containing a predetermined target population ofcells by (a) introducing a starting sample of cells into a growthmedium; (b) causing cells of the predetermined target population todivide; and (c) concurrently with, intermittently during, or followingstep (b), contacting the cells in the growth medium with a selectionelement, so as to select cells of the target population from other cellsin the growth medium; and (ii) administering the aliquot to the patient.

The selection element may include a plurality of selective bindingmolecules with affinity either for target cells or for a firstpopulation of non-target cells. The starting sample may be cord blood orbe derived from cord blood. The aliquot of cells may include CD 34+/−,Lin− cells. Expansion may be clonogenic.

The term “blood”, as used herein, refers to peripheral, fetal and cordblood, and is not meant to include bone marrow.

The phrase “blood-derived cells” refers to relatively undifferentiatedcells, and is not intended to include differentiated lymphoid cells suchas T or B cells.

Other features and advantages of the invention will be apparent from thedescription and drawings, and from the claims.

DESCRIPTION OF DRAWINGS

FIG. 1 is a diagram illustrating a method of treating a CNS-compromisedpatient according to one aspect of the invention.

FIGS. 2 and 3 illustrate methods of obtaining cells suitable for use inthe invention by selection. FIG. 2 is a highly enlarged diagrammaticview illustrating a method of positive selection of a target cell. FIG.3 is a highly enlarged diagrammatic view illustrating a method ofnegative selection of a non-target cell.

FIG. 4A is a schematic perspective view showing a rat that has beenprepared for a stroke model. FIG. 4B is a schematic diagram of anocclusion of the proximal MCA of the rat.

FIG. 5 is a schematic diagram showing a rat receiving anintraparenchymal administration of stem cells.

FIGS. 6-10 are graphs showing the results of behavioral tests performedon stroke model rats.

DETAILED DESCRIPTION

A preferred method of treating a human patient suffering from CNSimpairment or damage is shown schematically in FIG. 1. According to thismethod, the patient receives a therapeutic dose of blood-derived cells.The cells may be obtained, for example, from a bank of cord blood cells.Alternatively, cord blood cells can be put through a selectionprocedure, e.g., as described below and in U.S. Pat. No. 5,925,567, toselect for CD 34+/−, Lin− cells, and a relatively concentrated sample ofthese cells may be administered. If desired, the population of CD 34+/−,Lin− cells may be expanded prior to administration to the patient, asdescribed in U.S. Pat. No. 5,925,567. An adjuvant, e.g., a growth factoror genetic element, can be added to the cord blood cells or CD 34+/−,Lin− cells, or co-administered therewith, if desired.

After the cells are administered to the patient, e.g., using the methodsdescribed below, the CNS function of the patient is tested after aperiod of time, e.g., 90 days, to determine the extent of recovery offunction that has occurred. If recovery is determined to be inadequate,a further dose of cells will be administered and the evaluation processwill be repeated. If desired, CNS function can be tested morefrequently, e.g., after 30 days and, if treatment is repeated,periodically during such further treatment, in order to monitoreffectiveness of treatment and degree of improvement in CNS function.

The cells can be administered to a patient using any suitable technique.One suitable technique is intracerebral injection, either directly intothe brain (intraparenchymally) or into the spinal fluid(intraventricularly or intracisternally). The cells are generallycarried in a pharmaceutically acceptable liquid medium. Administrationcan be repeated or performed continuously (e.g., by a continuousinfusion into the spinal fluid). Multiple administrations are generallyspaced at least 2-7 days apart.

If the cells are to be injected into the brain, the patient's head isimmobilized in a standard stereotactic frame, and the site ofadministration of the cells is located, e.g., by standard CT or MRIscan. A small-bore hole is drilled in the skull, and the cells areinjected into the desired location using a syringe.

Suitable dosages of cells will vary depending on the amount of CNSdamage or deterioration that the patient has sustained, the weight ofthe patient, and other factors. Generally, the dosage may range fromabout 100,000 to 1,000,000,000 cells per administration, typically fromabout 1,000,000 to 10,000,000 cells.

Preferably, the cells administered are non-fetal, non-embryonic and arederived from blood. Suitable sources include fresh cord blood, CD 34+/−,Lin− cells separated from cord blood, and CD 34+/−, Lin− cells derivedby expanding cells selected from cord blood.

In the case of a selected and/or expanded population, the cells arepreferably CD 34+/−, Lin− cells or precursor cells that arecharacterized as: CD2⁻, CD3⁻, CD14⁻, CD16⁻, CD19⁻, CD24⁻, CD56⁻, CD66b⁻,glycophorin A⁻, flk-1⁺, CD45⁺, CXCR4⁺, MDR⁺.

Suitable procedures for separating CD 34+/−, Lin− cells from cord blood,and expanding the separated population, are described in U.S. Pat. No.5,925,567 and summarized below.

Separation/Expansion Procedures

FIGS. 2 and 3 illustrate suitable selection procedures. According tothese procedures, cells of a desired target population may besubstantially continuously proliferated by providing a system containinga nutrient medium in which cell proliferation can occur, and selectingcells of the target population from non-target cells in the system,concurrently with proliferation, intermittently during proliferation orfollowing proliferation. Cell proliferation and cell selection can becarried out using an almost infinite variety of different techniques andsettings, of which only a few are described below by way of example.Many other techniques will be readily perceived by those skilled in theart.

All of the preferred techniques, however, are based on the concepts ofpositive selection (providing a selection element having an affinity for(“selecting”) target cells), and negative selection (providing aselection element having an affinity for (“selecting”) non-targetcells). These two techniques used alone or in combination, allowunwanted cells to be removed from the system and target cells to beharvested whenever desired.

An example of a positive selection technique is illustrateddiagrammatically in FIG. 2. Briefly, one or more biotintylatedantibodies, specific for the target cells, and avidin are sequentiallyintroduced into the system. After a specified incubation time anybiotintylated antibody and avidin which have not formed a complex withthe target cells are rinsed away. Biotintylated dextran-iron is thenadded to the cell suspension. The biotintylated dextran-iron reacts withthe AvidinBiotintylated Antibody/Antigen Complex to form a largercomplex containing the biotintylated dextran-iron. This suspension isthen passed through a magnetic field. Positively selected cells remainin the magnetic field while cells which do not have the iron conjugatedcomplex are removed. After capture and rinsing, the magnetic field isremoved and the positively selected predetermined target population isreturned to the nutrient medium.

An example of a negative selection technique is illustrateddiagrammatically in FIG. 3. Briefly, one or more anti-dextran conjugatedantibodies specific for a predetermined population which is not of thepredetermined target population is introduced into the culture. After aspecified incubation time the cell suspension is passed through a columncontaining dextran coated glass beads. AnAntigen/Antibody/Anti-dextran/Dextran/Bead Complex forms, removing cellsnot of the predetermined target population from the nutrient medium. Thepredetermined target population is collected downstream and returned tothe nutrient medium.

Clearly, many other techniques could be utilized for both positive andnegative selection, as long as the desired affinity is provided by theselection element.

The selection element can be simply the selection molecule itself, orcan include other components, e.g., a solid support onto which theselection molecule is bound. The solid support can be formed of amaterial that will aid in performing the selection or in maintaining theselection molecules in a desired position or introducing and removingthem from the system. For example, as described above with reference toFIG. 2, the selection molecule can be bound to iron or other magneticparticles to allow the selected cells to be easily removed from thesystem by application of a magnetic field and then collected by removalof the magnetic field. Alternatively, the selection molecules can bebound onto the wall of a vessel containing the nutrient medium, or of achamber through which the nutrient medium flows during the method. Glassor other inert, impermeable beads can also be used as a solid support,as will be discussed in detail below. If beads or other particles areused, they can be provided in a packed configuration, through which thenutrient medium flows, or can be introduced into the system in a looseform, suspension, or in any desired type of array. As will be readilyunderstood, a wide variety of other solid supports can be used.

Adjuvants

Various adjuvants may be administered to the patient to further enhancerecovery of CNS function. The aliquot of cells may include variousadjuvants, or adjuvants may be co-administered with the cells. Suitableadjuvants include neural stimulants such as growth factors andneurotransmitters. Genetic elements may also be used as adjuvants, aswill be discussed below.

Suitable growth factors are discussed in co-pending application U.S.Ser. No. 60/149,561, filed Aug. 18, 2000, the disclosure of which isincorporated by reference herein. Suitable growth factors includemembers of the fibroblast growth factor (FGF) family, e.g., basic FGF,acid FGF, the hst/Kfgf gene product, FGF-5, or int-2; the neurotrophinfamily, e.g., nerve growth factor (NGF), brain-derived neurotrophicfactor (BDNF), neurotrophin 3 (NT3), or neurotrophin 4/5 (NT4/5); theinsulin-like growth factor family, including IGF-1 and IGF-2; theciliary neurotrophic growth factor (CNTF) family; the EGF family,including EGF, TGF-alpha, and HB-EGF; the TGF-beta family, includingTGFbeta and members of the BMP subfamily; leukemia inhibitory factor(LIF); oncostatin M; interleukins such as IL-II; members of theplatelet-derived growth factor (PDGF) family; and the VEGF family.Bioactive fragments, analogues and active fragments of theabove-mentioned growth factors may also be used.

The preferred dosage of the growth factor(s) will depend upon the resultdesired and the route of administration. The growth factor(s) may beadministered with the cells, or by a different route, including directlyinto the brain (intracerebrally), systemically (e.g., intravenously) orinto the spinal fluid (intracisternally or intracerebroventricularly).If administered directly into the brain or into the spinal fluid, thepreferred total amount of growth factor per administration is generallyfrom about 0.001 to 1000 mg, preferably about 0.01 to 100 mg. Ifadministration is systemic, the dosage will generally be based on thepatient's body weight, with typical dosages being from about 0.001 to100 mg/kg, preferably about 0.01 to 10 mg/kg.

Suitable neurotransmitters include neurotransmitter agonists orantagonists such as Prozac, amphetamines, Ritalin, and tricyclicantidepressants such as Elavil. These compounds act as neuralstimulants, and have well-established effects on the brain.

Administration of a genetic element may enhance recovery of a patientfrom a genetic disease such as Tay-Sachs. For example, if a patient issuffering from a disease resulting from the lack of function ordysfunction of a naturally occurring protein, as a result of a genemutation or genetic flaw, an exogenous gene may be installed on thecells to be administered to the patient. This is generally necessarywhen the cells that are administered are autologous cells. If allogeneiccells, e.g., immunologically matched cells, are administered to thepatient, it is generally not necessary to install an exogenous gene, asthe necessary gene would typically be present endogenously and wouldfunction properly. However, in some cases it may be desirable to installan exogenous gene in this case as well.

Other adjuvants may be administered, as discussed in co-pendingapplication U.S. Ser. No. 60/149,561, e.g., cell differentiation factorssuch as retinoic acid, neural guidance molecules such as semaphorins,netrins, neuropilins and ephrins, and physical treatments such astranscranial magnetic stimulation.

Evaluating CNS Function

Advantageously, the methods of the invention generally result in ameasurable improvement in CNS function, e.g., measurable strokerecovery. Tests that may be used to evaluate stroke recovery in humansinclude global functional and neurological outcome scores (ModifiedRankin Scale, NIH stroke scale, and Barthel Index); specialized tests ofneurological function (Fugl-Meyer Scale of motor functioning;neuropsychological test battery) and other tests, e.g., the testsdescribed in U.S. Pat. No. 5,885,231, the disclosure of which isincorporated by reference herein. Other tests that may be used toevaluate CNS function are well known in the neurological field.

EXAMPLE

Stem cells (CD 34+/−, Lin− cells) were selected from a sample of freshcord blood cells using the procedure described in Example 5 of U.S. Pat.No. 5,925,567, incorporated by reference above.

20 male Sprague Dawley rats, each weighing 300-350 grams, wereanesthetized with a 2% halothane with nitrous oxide/oxygen mixture(2:1), and subjected to a MCA occlusion, using a Modified Tamura model(see FIGS. 4A and 4B). This stroke model has been described in theliterature (see, e.g., Kawamata, T., et al., Intracisternal BasicFibroblast Growth Factor (bFGF) Enhances Functional Recovery andUpregulates the Expression of a Molecular Marker of Neuronal SproutingFollowing Focal Cerebral Infarction. Proc. Natl. Acad. Sci., 1997.94: p.8179-8184).

The rats received cefazolin sodium i.p. (40 mg/kg) one day beforesurgery and immediately following surgery. 24 hours after the occlusion,all of the rats received an injection directly into the brain tissuesurrounding the stroke (FIG. 5). 10 of the rats were injected with1,000,000 stem cells each; the other 10 rats were injected withphysiological buffered saline (PBS) vehicle. After injection, the ratswere given cyclosporin i.p. each day (10 mg/kg),

The rats were tested 3 days after the occlusion surgery, and then every4 days thereafter for the next 21 days to determine their centralnervous system function. The rats had also been tested, using the sametests, one day prior to the occlusion surgery (day −1). They were testedusing several behavioral tests, described below. For these tests, ratsare handled for 10 min. each day for seven days before stroke surgery.

The forelimb and hindlimb placing tests, described briefly below, aredescribed in further detail in Kawamata et al., supra. For the forelimbplacing test, the examiner holds the rat close to a table top and scoresthe rat's ability to place the forelimb on the table top in response towhisker, visual, tactile, or proprioceptive stimulation. Similarly, forthe hindlimb placing test, the examiner assesses the rat's ability toplace the hindlimb on the table top in response to tactile andproprioceptive stimulation. Separate subscores are obtained for eachmode of sensory input and added to give total scores (for the forelimbplacing test: 0=normal, 12=maximally impaired; for the hindlimb placingtest: 0=normal; 6=maximally impaired).

The body swing test (described in detail in Borlongan et al.,Transplantation of Cryopreserved Human Embryonal Carcinoma-derivedNeurons (NT2N Cells) Promotes Functional Recovery in Ischemic Rats, Exp.Neurol., 1998. 149: p. 310-321) has been used to examine sidepreferences after stroke. The animal is held approximately 1 inch fromthe base of its tail. It is then elevated to an inch above a surface ofa table. The animal is held in the vertical axis, defined as no morethan 10° to either the left or the right side. A swing is recordedwhenever the animal moves its head out of the vertical axis to eitherside. Before attempting another swing, the animal must return to thevertical position for the next swing to be counted. Thirty total swingsare counted. A normal animal typically has an equal number of swings toeither side. Following focal stroke, the animal tends to swing to thecontralateral side. There is a slow spontaneous recovery of body swingduring the first month after stroke.

Two other tests were conducted, the cylinder test and the paw reachingtest. The cylinder, or “spontaneous limb” test (described in detail inJones, T. A. & Schallert, T., Use-dependent Growth of Pyramidal NeuronsAfter Neocortical Damage, J. Neurosci., 1994. 14: p. 2140-2152),examines the forelimb side preference of animals as they rear up in anarrow glass cylinder. Animals are placed in a narrow Plexiglas cylinder(18×30 cm) and videotaped for 5 min (or for a minimum number ofmovements) on the day before stroke surgery and at weekly intervalsthereafter. Videotapes are then scored for the number of spontaneousmovements made by each forelimb to initiate rearing, to land on or tomove laterally along the wall of the cylinder, or to land on the floorafter rearing. The mean number of spontaneous movements of each forelimbis then expressed as an asymmetry score (total contralateral forelimbuse−total ipsilateral forelimb use)/total forelimb use). Before stroke,animals tend to use both forelimbs equally. After stroke, there is apreference for the unimpaired (ipsilateral) forelimb. This asymmetryrecovers spontaneously to a partial degree during the first month afterstroke.

The paw reaching test (described in detail in Kolb et al., Nerve GrowthFactor Treatment Prevents Dendritic Atrophy and Promotes Recovery ofFunction After Cortical Injury, Neuroscience, 1997.76: p. 1139-1151),measures the dexterity of the forepaws in reaching through the bars of acage to grab and eat food pellets. Animals are food-deprived (allowing15 g regular food per day) before the training and testing dates. Theanimals are placed in the test cages (23×30×13 cm) with floors and wallconstructed of 2 mm bars, 9 mm apart edge to edge. A 8 cm wide and 6 crndeep tray, containing chocolate flavored pellets (45 mg/each), ismounted on one wall of the cage. The rats are required to extend aforelimb through the gap in the bars, grasp and retract the food. Thetray is 0.5 crn apart from the cage so that the rats cannot scrape thefood into the cage. Rats are trained for 10 days using both forepawsbefore stroke surgery. After stroke surgery, they are trained again for5 days using only the impaired (contralateral) paw. This is done byplacing a bracelet on the intact paw, which does not allow it to reachthrough the bars of the cage. Then, animals are videotaped for 5 min.with the bracelet on, and 5 min. with the bracelet off. The total numberof “reaches” and “hits” (successful grabbing and eating of food pellet)are recorded for each interval.

The results of these behavioral tests are shown in FIGS. 6-10. Theasterisks in FIGS. 6 and 7 indicate that data in the stem cell groupswere different from the vehicle groups by p<0.05 by two-way ANOV A(treatment X time). The lack of asterisks in FIGS. 8-10 indicate thatthere were no significant differences.

A statistically significant difference was observed between the ratsthat received stem cells and the rats that received the vehicle in theforelimb and hindlimb placing tests, with the rats that received stemcells showing significantly better recovery of function. For theforelimb placing test, all but one stem cell treated animal (9/10) didbetter than the mean of vehicle-treated animals by the end of theexperiment. For the hindlimb placing test, all stem cell treated animals(10/10) did better than the mean of vehicle-treated animals by the endof the experiment.

No significant difference was observed in the swinging, cylinder, or pawreaching tests.

While the data show mixed results, sufficient improvement was observedin the forelimb and hindlimb placing tests to lead the inventors tobelieve that the invention is viable. The tests performed measuredifferent aspects of sensorimotor recovery, and the forelimb andhindlimb placing tests may be the most sensitive to treatment effects.The inventors believe that the results of the other tests could perhapsbe improved by administering a higher dosage of cells or by repeatedadministration; these tests have not been conducted to date.

Other embodiments are within the claims. For example, the cells may bederived from other blood-based, non-fetal and non-embryonic sources.

1. A method for treating a human patient suffering from a disease ordisorder selected from multiple sclerosis, Tay-Sacks, and cerebralpalsy, said method comprising administering to said patient acomposition comprising human CD34+/−, Lin− cells from human umbilicalcord blood (UCB) or peripheral blood, wherein said administering resultsin a measurable improvement in said disease or disorder in said patient.2. The method of claim 1, wherein said patient is administered whole UCBor peripheral blood.
 3. The method of claim 1, wherein said cells areisolated from UCB or peripheral blood.
 4. The method of claim 1, whereinsaid CD34+/−, Lin− cells are separated from other mononuclear cellspresent in said UCB or said peripheral blood prior to saidadministering.
 5. The method of claim 1 further comprising concurrentlywith or following administration of said CD34+/−, Lin− cellsadministering a growth factor to said patient.
 6. The method of claim 5,wherein said growth factor is selected from the group consisting ofoncostatin M, FGF, neurotrophin, IGF, CNTF, EGF, TGF-beta, LIF,interleukins, PDGF, and VEGF.
 7. The method of claim 1, wherein saidCD34+/−, Lin− cells are allogeneic cells or autologous cells.
 8. Themethod of claim 1, wherein said CD34+/−, Lin− cells are characterized asnegative for expression of CD2, CD3, CD14, CD16, CD19, CD24, CD56,CD66b, and glycophorin A, and positive for expression of flk-1, CD45,CXCR4, and MDR.
 9. The method of claim 1, wherein said method comprisesadministering a population of cells consisting of human CD34+/−, Lin−cells.
 10. The method of claim 1, wherein said UCB or peripheral bloodis administered to said patient systemically, intercerebrally,intracisternally, intracerebroventricularly, or intraparenchymally. 11.A method for treating a human patient suffering from a disease ordisorder selected from stroke, Parkinson's Disease, Alzheimer's Disease,Huntington's Disease, amyotrophic lateral sclerosis (ALS), multiplesclerosis, Tay-Sacks, and cerebral palsy, said method comprisingadministering to said patient a composition comprising human CD34+/−,Lin− cells from human umbilical cord blood (UCB) or peripheral blood,wherein said CD34+/−, Lin− cells are separated from other mononuclearcells present in said UCB or said peripheral blood prior to saidadministering and wherein said administering results in a measurableimprovement in said disease or disorder in said patient.
 12. The methodof claim 11 further comprising concurrently with or followingadministration of said CD34+/−, Lin− cells administering a growth factorto said patient.
 13. The method of claim 12, wherein said growth factoris selected from the group consisting of oncostatin M, FGF,neurotrophin, IGF, CNTF, EGF, TGF-beta, LIF, interleukins, PDGF, andVEGF.
 14. The method of claim 11, wherein said CD34+/−, Lin− cells areallogeneic cells or autologous cells.
 15. The method of claim 11,wherein said CD34+/−, Lin− cells are characterized as negative forexpression of CD2, CD3, CD14, CD16, CD19, CD24, CD56, CD66b, andglycophorin A, and positive for expression of flk-1, CD45, CXCR4, andMDR.
 16. The method of claim 11, wherein said method comprisesadministering a population of cells consisting of human CD34+/−, Lin−cells.
 17. The method of claim 11, wherein said UCB or peripheral bloodis administered to said patient systemically, intercerebrally,intracisternally, intracerebroventricularly, or intraparenchymally.